This is a proposal for Cedars-Sinai Medical Center to become a Discovery Site for the MAPP Network. We have assembled an exceptional multidisciplinary team, which includes over 20 years of experience in NIH-funded urology research center leadership; a long track record of discovery and hypothesis-driven research and NIH funding in benign urologic conditions; an NIH-funded track record in studies of human subjects; a distinguished record in clinical biomarker discovery based on implementation of state-of-the-art mass spectrometry-based proteomics and related technologies; leadership in research and discovery of complex microbial communities; and translational pathology and biobanking. Cedars-Sinai Medical Center (CSMC) is one of the largest academic medical centers in the western US. Our overall objective is to use state-of-the-art approaches to develop novel strategies for phenotyping and ultimately improving the clinical care of patients with urologic chronic pelvic pain syndromes (UCPPS). The overall hypothesis of this project is that uncultivated commensal microbial communities and their interactions with the host are associated with the development of UCPPS and that the resultant protein patterns in the urine and blood create a signature diagnostic of UCPPS. We will use two complementary approaches to test this hypothesis: (1) We will employ state-of-the-art resources in microbiome genomic sequencing and characterization, some of which are unique to Cedars-Sinai, to define the microbiome/mycobiome of UCPPS patients in comparison to normal subjects, (2) We hypothesize that UCPPS is caused by the changes of some proteins at the expression and post-translational modification levels and that these changes can be rapidly identified in an unbiased manner using advanced proteomics technologies. The Specific Aims are: Aim 1. To identify disease-specific changes in bladder commensal bacterial and fungal communities by next generation sequencing of ribosomal DNA in urine from MAPP patients with UCPPS and control subjects. Aim 2: To identify clinically relevant non-invasive urinary biomarkers of UCPPS through deep proteomics analysis of urine and blood from MAPP patients. Results from the CSMC team and the multidisciplinary interactions promoted by this proposal will lay the groundwork for innovative collaborative studies on UCPPS within the MAPP network.